1. FIELD OF THE INVENTION
The present invention relates to a hydrophilic porous spherical separating carrier and its preparation. More particularly, it relates to a hydrophilic separating carrier suitable for separating important water soluble materials such as proteins and enzymes in an aqueous medium by a gel permeable effect in a high speed liquid chromatography.
2. DESCRIPTION OF THE PRIOR ARTS
Hertofore, a gel filtering method has been known as one important method for separating and purifying water soluble materials.
Dextran gel and polyacrylamide gel have been widely used in a biochemical field etc., as a separating carrier. However, these gels have low mechanical strength whereby these gels could not be used as carriers for high speed liquid chromatography, disadvantageously.
Recently, various tests for producing hydrophilic gels having high mechanical strength by a crosslink polymerization of an acrylate monomer or a methacrylate monomer, have been studied. However, it has not been attained to obtain a gel having high function for satisfying the following important functions.
(1) High mechanical strength PA1 (2) No adsorption of measured materials PA1 (3) High separation factor (resolution) PA1 Apparatus: HLC 802 UR (manufactured by Toyo Soda Ind. Co.) PA1 Column: TSK GEL G-2000H inner diameter 7.5 mm.times.length 600 mm two columns PA1 Eluent: tetrahydrofuran PA1 Flow rate: 1 ml/min. PA1 Temperature in measurement: room temperature PA1 Detector: Differential refractometer
These functions can be obtained only when various desired physical and chemical properties required for an ideal gel are given.
Among the known gels, polyethyleneglycol dimethacrylate gels disclosed in Japanese Patent Publication No. 24512/1973 have an advantage of small adsorbing property but have disadvantages of low mechanical strength and small separation factor.
The gels obtained by copolymerizing hydroxyethyl methacrylate and ethyleneglycold methacrylate disclosed in Japanese Unexamined Patent Publication No. 64187/1973 have high adsorbing property and low mechanical strength.
The inventors have studied on various crosslinking agents in view of the consideration that the most important matter for producting ideal hydrophilic separating carriers is in a selection of a crosslinking agent.
As result, it has been found that the conventional crosslinking agent such as ethyleneglycoldimethacrylate is effective for increasing mechanical strength of a gel but has inferior hydrophilic property to give strong hydrophobic adsorption sites in the gel on the properties required for the crosslinked product.